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Viruses 2016, 8(11), 306; doi:10.3390/v8110306

Epitope Identification and Application for Diagnosis of Duck Tembusu Virus Infections in Ducks

1
State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute of Chinese Academy of Agricultural Sciences, Harbin 150001, China
2
School of Electrical Engineering & Automation, Harbin Institute of Technology, Harbin 150006, China
3
Institute of Genomics and Bioinformatics, National Chung Hsing University, Taichung 402, Taiwan
These authors contributed equally to this work.
*
Authors to whom correspondence should be addressed.
Academic Editor: Michael R. Holbrook
Received: 3 August 2016 / Revised: 4 November 2016 / Accepted: 4 November 2016 / Published: 10 November 2016
(This article belongs to the Special Issue Advances in Flavivirus Research)
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Abstract

Duck Tembusu virus (DTMUV) causes substantial egg drop disease. DTMUV was first identified in China and rapidly spread to Malaysia and Thailand. The antigenicity of the DTMUV E protein has not yet been characterized. Here, we investigated antigenic sites on the E protein using the non-neutralizing monoclonal antibodies (mAbs) 1F3 and 1A5. Two minimal epitopes were mapped to 221LD/NLPW225 and 87YAEYI91 by using phage display and mutagenesis. DTMUV-positive duck sera reacted with the epitopes, thus indicating the importance of the minimal amino acids of the epitopes for antibody-epitope binding. The performance of the dot blotting assay with the corresponding positive sera indicated that YAEYI was DTMUV type-specific, whereas 221LD/NLPW225 was a cross-reactive epitope for West Nile virus (WNV), dengue virus (DENV), and Japanese encephalitis virus (JEV) and corresponded to conserved and variable amino acid sequences among these strains. The structure model of the E protein revealed that YAEYI and LD/NLPW were located on domain (D) II, which confirmed that DII might contain a type-specific non-neutralizing epitope. The YAEYI epitope-based antigen demonstrated its diagnostic potential by reacting with high specificity to serum samples obtained from DTMUV-infected ducks. Based on these observations, a YAEYI-based serological test could be used for DTMUV surveillance and could differentiate DTMUV infections from JEV or WNV infections. These findings provide new insights into the organization of epitopes on flavivirus E proteins that might be valuable for the development of epitope-based serological diagnostic tests for DTMUV. View Full-Text
Keywords: duck Tembusu virus; E protein epitopes; type specific and cross-reactive epitopes; E protein 3D structure; diagnosis duck Tembusu virus; E protein epitopes; type specific and cross-reactive epitopes; E protein 3D structure; diagnosis
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MDPI and ACS Style

Li, C.; Liu, J.; Shaozhou, W.; Bai, X.; Zhang, Q.; Hua, R.; Liu, J.-H.; Liu, M.; Zhang, Y. Epitope Identification and Application for Diagnosis of Duck Tembusu Virus Infections in Ducks. Viruses 2016, 8, 306.

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